Human umbilical cord mesenchymal stem cell exosomes deliver potent oncolytic reovirus to acute myeloid leukemia cells.

In addition to chemotherapy, oncolytic viruses are an efficient treatment for acute myeloid leukemia (AML). Like other oncolytic viruses, the anti-tumor efficacy of reovirus when administered intravenously is reduced due to the presence of neutralizing antibodies. In this study, we evaluated the role of exosomes in human umbilical cord-derived mesenchymal stem cells (UC-MSCs) to deliver reovirus to AML cells. We show that UC-MSCs loaded with reovirus can deliver reovirus to tumor cells without cellular contact. We further demonstrate that the exosome inhibitor, GW4869, inhibits the release of exosomes as well as inhibited the transfer of reovirus from UC-MSCs to tumor cells. Mechanistically, we show that exosomes derived from reovirus-infected UC-MSCs (MSCREO-EXOs) have a tumor lysis effect and transmit reovirus to tumor cells mainly through clathrin-mediated endocytosis (CME) and macropinocytosis. In addition, we demonstrate the feasibility of using MSC-derived exosomes (MSC-EXOs) as a reovirus carrier to exert an anti-tumor effect on AML cells. Collectively, our data indicate that UC-MSCs transfer reovirus to AML cells via exosome release and prompt further study of MSC-EXOs as a potential reovirus carrier to treat AML.

Large-scale Isolation of Exosomes Derived from NK Cells for Anti-tumor Therapy.

Exosomes are lipid bilayer-enclosed vesicles, actively secreted by cells, containing proteins, lipids, nucleic acids, and other substances with multiple biological functions after entering target cells. Exosomes derived from NK cells have been shown to have certain anti-tumor effects and potential applications as chemotherapy drug carriers. These developments have resulted in high demand for exosomes. Although there has been large-scale industrial preparation of exosomes, they are only for generally engineered cells such as HEK 293T. The large-scale preparation of specific cellular exosomes is still a major problem in laboratory studies. Therefore, in this study, we used tangential flow filtration (TFF) to concentrate the culture supernatants isolated from NK cells and isolated NK cell-derived exosomes (NK-Exo) by ultracentrifugation. Through a series of characterization and functional verification of NK-Exo, the characterization, phenotype, and anti-tumor activity of NK-Exo were verified. Our study provides a considerably time- and labor-saving protocol for the isolation of NK-Exo.

NK cell-derived exosomes enhance the anti-tumor effects against ovarian cancer by delivering cisplatin and reactivating NK cell functions.

Exosomes are membranous vesicles actively secreted by almost all cells and they deliver certain intracellular molecules, including nucleic acids, proteins, and lipids, to target cells. They are also considered to be good carriers for drug delivery due to their biocompatibility, high permeability, low immunogenicity, and low toxicity. Exosomes from immune cells were also reported to have immunomodulatory activities. Herein we evaluated the application of exosomes derived from expanded natural killer cells (eNK-EXO) for the treatment of ovarian cancer (OC). We demonstrate that eNK-EXO express typical protein markers of natural killer (NK) cells, can be preferentially uptaken by SKOV3 cells, and display cytotoxicity against OC cells. Furthermore, eNK-EXO loaded with cisplatin could sensitize drug-resistant OC cells to the anti-proliferation effect of cisplatin. In addition, we show that eNK-EXO could activate NK cells from immunosuppressive tumor microenvironment, the mechanism of which is explored by transcriptional analysis. In summary, eNK-EXO exhibit anti-tumor activity against OC on its own, could be used to deliver cisplatin and enhance its cytotoxic effect against drug-resistant OC cells and also reverse the immunosuppression of NK cells, which may lead to great prospect of using eNK-EXO in the treatment of OC in the clinic. Our work also builds a strong foundation for further evaluation of eNK-EXO in other solid tumor therapies.

Correlation of VEGF and EGFR in peripheral blood with clinical stage and pathological grade of renal cell carcinoma and analysis of prognosis.

PURPOSE: To detect the expression of VEGF and EGFR in peripheral blood and cancer tissues of patients with renal cell carcinoma (RCC), and to explore the correlations with clinical stage, pathological grade and prognosis of disease. METHODS: A total of 64 patients with RCC who were diagnosed and treated from June 2016 to August 2017 in our hospital were enrolled. Patients were divided into different clinical stages and pathological grades, and ELISA and immunohistochemistry were used to detect the expression of VEGF and EGFR in peripheral blood. Peripheral blood was also taken from 24 healthy individuals to serve as control group. Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the expression of VEGF and EGFR in RCC tissues and paracancer tissues. All patients were followed up after discharge to record their survival. RESULTS: Significant differences in the expression levels of VEGF and EGFR were found between stage III and IV (p<0.05), but not between stage I and II. Expressions level of VEGF and EGFR in serum of well-differentiated, moderatelydifferentiated, and poorly-differentiated RCC were all higher than those in the healthy control group, and significant differences were found between different pathological grades (p<0.05). Patients with higher expression levels of VEGF and EGFR showed shorter survival compared to patients with lower expression levels (p<0.05). CONCLUSION: VEGF and EGFR in peripheral blood can be used as one of the effective indicators of prognosis of RCC. Our study provided reference for clinical treatment and prediction of prognosis of RCC.

Identification and Validation of Gene Expression Patterns in Cystitis Glandularis Patients and Controls.

Our aim was to investigate differences in gene expression in bladder tissues between cystitis glandularis (CG) patients and healthy controls. Subsequent RNA was isolated from urinary bladder samples from CG patients and healthy controls, followed by RNA sequencing analysis. There were 4263 differentially expressed genes in urinary bladder between CG patients and controls, and 8 genes were verified with real-time PCR, Western blot, and enzyme-linked immunosorbent assay (ELISA) analysis. Gene set enrichment analysis (GSEA) revealed that 25 signaling pathways were upregulated in CG patients, and 17 signaling pathways were found upregulated in healthy controls. The mRNA expression levels of the indicated genes, including CCND1, CCNA1, EGFR, AR, CX3CL1, CXCL6, and CXCL1, were significantly increased in urinary bladder from CG and bladder cancer (BC) patients compared with healthy controls, while TP53 was decreased. CX3CL1, CXCL6, and CXCL1 concentrations in peripheral blood from CG and BC patients were significantly increased compared with healthy controls. The protein expression levels of CCND1, EGFR, and AR were significantly increased in urinary bladder from CG and BC patients compared with healthy controls. In conclusion, the gene expression profile of CG patients has established a foundation to study the gene mechanism of CG and BC progression.

Transcriptome analysis of primary aldosteronism in adrenal glands and controls.

Primary aldosteronism (PA) is the most common form of endocrine hypertension. This study was to investigate the gene expression profile in PA adrenal glands and normal controls using RNA-Sequencing. By performing transcriptome analyses for 3 PA adrenal glands and 3 controls on Illumina platform, we identified 1,093 transcripts as significantly differently expressed genes (DEGs), which provided clues for further study of these transcript changes during PA pathogenesis. Further, Gene Set Enrichment Analysis (GSEA) identified 35 significant Kyoto Encyclopedia of Genes and Genomes (KEGG) biological pathways, including 'ribosome', 'oxidative phosphorylation', 'histidine metabolism', 'xenobiotics metabolism by Cytochrome P450', 'drug metabolism by Cytochrome P450', 'tyrosine metabolism' and 'glutathione metabolism'. In summary, we identified novel genes that are associated with PA phenotype, as well as differently regulated biological pathways relating to protein synthesis, energy acquisition and metabolism. Our study provides new candidates for further elucidation of the molecular mechanisms underlying PA pathogenesis.